Stabilizing fALS SOD1 Variants by Crosslinking Subunits

通过交联亚基稳定 fALS SOD1 变体

• 批准号: 8071048
• 负责人: Jeffrey Neil Agar
• 金额: $ 19.36万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8071048
• 关键词: Amino Acids; Amyotrophic Lateral Sclerosis; Binding; Biochemical Reaction; Biological Assay; Bridged Compounds; Chemicals; Coupling; Cuprozinc Superoxide Dismutase; Cyclic Peptides; Cysteine; Data; Disease; Dissociation; Disulfides; FDA approved; Familial Amyotrophic Lateral Sclerosis; Generations; Genes; Glutathione; Goals; In Vitro; Individual; Laboratories; Length; Libraries; Maleimides; Mass Spectrum Analysis; Mediating; Methods; Modeling; Molecular Chaperones; Mutation; Neurodegenerative Disorders; Pathway interactions; Patients; Peptides; Performance; Peroxides; Pharmaceutical Preparations; Physiological; Positioning Attribute; Property; Proteins; Proxy; Quality of life; Quantitative Structure-Activity Relationship; Reaction; Reducing Agents; Reporting; Research Personnel; Services; Side; Site; Specificity; Structure; Sulfhydryl Compounds; Sulfur; Surface; Testing; Therapeutic; Toxic effect; Variant; Work; arm; base; chemical reaction; combinatorial; complement C2a; crosslink; cysteinylcysteine; data exchange; dimer; disulfide bond; gain of function; improved; in vivo; interest; melting; monomer; novel; oxidation; prevent; public health relevance; small molecule

DESCRIPTION (provided by applicant): Over 120 mutations in the gene encoding SOD1 are associated with amyotrophic lateral sclerosis. SOD1 mutations result in a toxic property, and a prevailing hypothesis is that a gain-of-function involves dissociation of the SOD1 dimer followed by aggregation. As a result, stabilization of the SOD1 dimer has become an approach for SOD1 therapy. We have discovered a specific and stoichiometric method for stabilizing the SOD1 dimer by tethering Cys111 residues with a covalent cross-linker. Cys111 is a surface residue situated at the dimer interface, and only a short distance apart from the Cys111 (8¿) on an opposing monomer. Our hypothesis is that compounds that bridge Cys111 residues will function as novel pharmacological chaperones that stabilize fALS SOD1 variants and that peptide-based cross-linkers are suitable for developing a quantitative structure activity model that can be used to create a second generation of improved peptides. In the hopes of inspiring interest, our first aim is to expand the scope of our original findings to include the five most prevalent fALS variants. While our first generation cross-linkers work well in vitro, they known to be toxic in vivo. Therefore, our second aim is to create a new class of peptide-based chemicals that are optimized to cross-link Cys111. PUBLIC HEALTH RELEVANCE: The neurodegenerative diseases, including ALS, have proven extraordinarily difficult to treat, for example, ALS has only one FDA-approved drug for its treatment, reporting little or no effect on patient survival and quality of life. Until recently researchers did not understand the mechanism of ALS- associated proteins' toxicity, but now many believe this involves the dissociation of the protein SOD1 from a dimer into two monomers. Preventing this dissociation has become a focus for therapy, and we have discovered a novel and effective in vitro strategy for stabilizing SOD1 that cross-links individual monomers. This proposal aims to expand the scope of our original findings by testing the most prevalent SOD1 variants, and to develop and optimize molecules that are less toxic than our first generation of cross-linkers.

描述（由申请人提供）：编码SOD 1的基因中超过120个突变与肌萎缩侧索硬化症相关。SOD 1突变导致毒性，普遍的假设是功能获得涉及SOD 1二聚体的解离，然后聚集。因此，SOD 1二聚体的稳定化已成为SOD 1治疗的方法。我们已经发现了一种特定的和化学计量的方法，用于稳定的SOD 1二聚体通过栓系Cys 111残基与共价交联剂。Cys 111是位于二聚体界面处的表面残基，并且与相对单体上的Cys 111（8 <$）仅相距很短的距离。我们的假设是，桥接Cys 111残基的化合物将作为稳定fALS SOD 1变体的新型药理学伴侣，并且基于肽的交联剂适合于开发可用于创建第二代改进肽的定量结构活性模型。为了激发人们的兴趣，我们的第一个目标是扩大我们原始发现的范围，包括五种最常见的fALS变体。虽然我们的第一代交联剂在体外工作良好，但已知它们在体内有毒。因此，我们的第二个目标是创建一类新的基于肽的化学品，这些化学品被优化以交联Cys 111。 公共卫生相关性：包括ALS在内的神经退行性疾病已被证明非常难以治疗，例如，ALS只有一种FDA批准的药物用于治疗，报告对患者生存和生活质量几乎没有影响。直到最近，研究人员还不了解ALS相关蛋白毒性的机制，但现在许多人认为这涉及蛋白质SOD 1从二聚体解离成两个单体。防止这种解离已成为治疗的焦点，我们已经发现了一种新的和有效的体外策略，用于稳定SOD 1交联单个单体。该提案旨在通过测试最常见的SOD 1变体来扩大我们原始发现的范围，并开发和优化比我们第一代交联剂毒性更低的分子。